Transgenic synaptotagmin is expressed at appropriate levels and exhibits appropriate localization.

<p>(A-C) Representative western blots above and average synaptotagmin expression normalized to actin levels below. <i>Syt</i> heterozygotes expressing one copy of the mutant transgene (<i>+/-;P[syt^P-L]/+</i>, n = 6, 5, and 7, respectively) are compared to <i>syt</i> heterozygotes (<i>+/-</i>, n = 5, A), <i>syt</i> homozygotes (<i>+/+</i>, n = 5, B), and <i>syt</i> heterozygotes expressing one copy of the wild type transgene (<i>+/-; P[syt^WT]/+</i>, n = 8, C). The <i>P[syt^P-L]</i> heterozygotes had approximately twice as much synaptotagmin expression as <i>syt</i> heterozygotes (p = 0.0008, student t-test), while there was no significant difference in expression between the other lines compared (Fig 2B, p = 0.69 and Fig 2C, p = 0.25). Error bars depict SEM. (D, E) Third instar larvae stained with antibodies against horseradish peroxidase (HRP) as a general axonal stain (red) and synaptotagmin (green). Scale bars represent 10 μm. (F, G) First instar larvae stained with antibodies against synaptotagmin. Scale bars represent 20 μm.